11,371 results match your criteria Journal of Immunological Methods [Journal]


A dual neutrophil-T cell purification procedure and methodological considerations in studying the effects of estrogen on human Th17 cell differentiation.

J Immunol Methods 2019 Feb 13. Epub 2019 Feb 13.

Program in Translational Medicine, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children Research Institute, Canada; Departments of Paediatrics and Physiology, University of Toronto, Canada; Institute of Medical Sciences, University of Toronto, Canada. Electronic address:

New procedures are required to optimize the use of blood samples to study different cell types. The purification of neutrophils and T cells from the same blood sample is not commonly described. We have previously used PolymorphPrep™ (P) or LymphoPrep™ (L) for purifying neutrophils or T cells, respectively. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.002DOI Listing
February 2019

The enzymatic removal of immunoglobulin variable domain glycans by different glycosidases.

J Immunol Methods 2019 Feb 8. Epub 2019 Feb 8.

Sanquin Research, Department of Immunopathology, Amsterdam, The Netherlands; Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

About 15% of immunoglobulin G (IgG) molecules contain glycans linked to the antigen-binding fragments (Fab arms) in addition to the glycans linked to the crystallizable fragment (Fc tail) of all IgGs. Fab glycosylation appears to be an important feature of antibodies, for example by influencing antigen binding and antibody stability. The reliable generation of antibodies that either have or lack Fab glycans would be very helpful to study the role of Fab glycans in more detail. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.005DOI Listing
February 2019

A monoclonal antibody targeted to the functional peptide of αB-crystallin inhibits the chaperone and anti-apoptotic activities.

J Immunol Methods 2019 Feb 6. Epub 2019 Feb 6.

Sue Anschutz-Rodgers Eye Center and Department of Ophthalmology, University of Colorado School of Medicine, United States; Skaggs School of Pharmacy and Pharmaceutical Sciences, Anschutz Medical Campus, Aurora, CO 80045, United States. Electronic address:

αB-Crystallin is a member of the small heat shock protein family. It is a molecular chaperone and an anti-apoptotic protein. Previous studies have shown that the peptide (DRFSVNLDVKHFSPEELKVKV, hereafter referred to as peptain-1) from the core domain of αB-crystallin exhibits both chaperone and anti-apoptotic properties similar to the parent protein. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.004DOI Listing
February 2019

An improved clonogenic culture method for thymic epithelial cells.

J Immunol Methods 2019 Feb 6. Epub 2019 Feb 6.

Department of Immunology and Cell Biology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan; Center for iPS Cell Research and Application (CiRA), Laboratory of Immunobiology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan. Electronic address:

A clonogenic assay system for thymic epithelial cells (TECs) is of crucial importance for identifying thymic epithelial stem and/or progenitor cells, evaluating their activities, and understanding the mechanisms of thymic involution. However, current systems are not sufficiently sensitive at detecting and quantifying TEC colonies from the adult thymus. Here, we optimized the culture condition to detect visible colonies from adult TECs by modifying our previous culture methods. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.003DOI Listing
February 2019
1 Read

A novel method to efficiently isolate medullary thymic epithelial cells from murine thymi based on UEA-1 MicroBeads.

J Immunol Methods 2019 Feb 5. Epub 2019 Feb 5.

AG Clinical Immunology, Institute of Immunology, Rostock University Medical Center, Rostock, Germany.

Objective: The central mechanism for establishing a self-tolerant and functional T cell repertoire includes the promiscuous expression of otherwise tissue-restricted proteins by medullary thymic epithelial cells (TEC). We here demonstrate a novel and highly efficient method for isolating this rare key cell type.

Methods: We combined the enrichment of medullary TEC via UEA-1 MicroBeads with the subsequent depletion of residual CD45 hematopoietic cells via specific size exclusion and compared our results to the standard Percoll enrichment method and isolation procedure via flow cytometric cell sorting. Read More

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http://dx.doi.org/10.1016/j.jim.2019.02.001DOI Listing
February 2019

Optimized protocols for studying the NLRP3 inflammasome and assessment of potential targets of CP-453,773 in undifferentiated THP1 cells.

J Immunol Methods 2019 Feb 5. Epub 2019 Feb 5.

Inflammation & Immunology Research Unit, Pfizer, Cambridge, MA 02139, United States. Electronic address:

The NLRP3 inflammasome is a complex multimeric signaling apparatus that regulates production of the pro-inflammatory cytokine IL-1β. To overcome both the variability among primary immune cells and the limitations of genetic manipulation of differentiated human or murine macrophages, we developed a simplified, reliable and relevant cell-based model for studying the NLRP3 inflammasome using the undifferentiated human myelomonocytic cell line THP1. Undifferentiated THP1 cells constitutively express NLRP3, and NLRP3 inflammasome activation occurred in response to canonical NLRP3 activation stimuli including nigericin, ATP, and urea crystals, culminating in pro-IL-1β cleavage, extracellular release of mature IL-1β, and pyroptosis. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183040
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http://dx.doi.org/10.1016/j.jim.2019.02.002DOI Listing
February 2019
6 Reads

Optimisation, harmonisation and standardisation of the direct mycobacterial growth inhibition assay using cryopreserved human peripheral blood mononuclear cells.

J Immunol Methods 2019 Jan 30. Epub 2019 Jan 30.

The Jenner Institute, University of Oxford, UK.

A major challenge to TB vaccine development is the lack of a validated immune correlate of protection. Mycobacterial growth inhibition assays (MGIAs) represent an unbiased measure of the ability to control mycobacterial growth in vitro. A successful MGIA could be applied to preclinical and clinical post-vaccination samples to aid in the selection of novel vaccine candidates at an early stage and provide a relevant measure of immunogenicity and protection. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.006DOI Listing
January 2019

Development and validation of HIV-1 Multiplex Serology.

J Immunol Methods 2019 Mar 18;466:47-51. Epub 2019 Jan 18.

Infections and Cancer Epidemiology, Infection, Inflammation and Cancer Research Program, German Cancer Research Center (DKFZ), Heidelberg, Germany; Faculty of Biosciences, Heidelberg University, Heidelberg, Germany. Electronic address:

By inducing immunosuppression in infected patients, human immunodeficiency virus-1 (HIV-1) generates a favorable environment for opportunistic infections and the development of several human cancers. In order to detect individual serum or plasma HIV-1 antibody status for epidemiological studies, high-throughput HIV-1 Multiplex Serology was developed. Seven HIV-1 antigens were recombinantly expressed in E. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183035
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http://dx.doi.org/10.1016/j.jim.2019.01.007DOI Listing
March 2019
3 Reads

A novel tandem repeat cloning technique for creation of multiple short peptide repeats to differentiate closely related antigens.

J Immunol Methods 2019 Jan 17. Epub 2019 Jan 17.

Microbiology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka 570011, India.

Antibody cross-reactivity is a problem often associated with closely related antigens. This study was aimed to develop a method enabling differentiation of closely related toxins based on antigen designing strategy. The method involves identification of disparate amino acids (AA) confined to target antigen in comparison with two or more closely related antigens, their assembly into a DNA oligomer and further cloning as six tandem repeats (TR) using restriction and ligation strategy into a desired vector and finally generation of antigen specific antibodies. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.008DOI Listing
January 2019
1 Read

Development and validation of different indirect ELISAs for MERS-CoV serological testing.

J Immunol Methods 2019 Mar 16;466:41-46. Epub 2019 Jan 16.

Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.

Since 2012, MERS-CoV has caused up to 2220 cases and 790 deaths in 27 countries with Saudi Arabia being the most affected country with ~83.1% of the cases and ~38.8% local death rate. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.005DOI Listing
March 2019
1 Read
2.005 Impact Factor

Reducing the risk of misdiagnosis of indirect ELISA by normalizing serum-specific background noise: The example of detecting anti-FGFR3 autoantibodies.

J Immunol Methods 2019 Mar 14;466:52-56. Epub 2019 Jan 14.

Synaptopathies and Autoantibodies, Faculty of Medicine Jacques Lisfranc, University Jean Monnet, University of Lyon, Saint-Étienne, France; Synaptopathies and Autoantibodies, Institut NeuroMyoGene INSERM U1217/CNRS UMR 5310, University of Lyon, Université Claude Bernard Lyon 1, Lyon, France; Neurology Department, Centre Hospitalier Universitaire de Saint-Étienne, Saint-Étienne, France.

Indirect enzyme-linked immunosorbent assay (ELISA) is an important diagnostic method as it enables the quantification of the presence of autoantibodies in human blood sera. However, unspecific binding of antibodies to the solid phase causes considerable serum-specific background noise (SSBN), involving the risk of false positive diagnosis. Therefore, we present a simple and concise, yet obvious proof-of-principle of a recently suggested normalization method. Read More

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http://dx.doi.org/10.1016/j.jim.2019.01.004DOI Listing
March 2019
1 Read

Determination of human γδ T cell-mediated cytotoxicity using a non-radioactive assay system.

J Immunol Methods 2019 Mar 14;466:32-40. Epub 2019 Jan 14.

Center for Bioinformatics and Molecular Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Program for Nurturing Global Leaders in Tropical and Emerging Infectious Diseases, Graduate School of Biomedical Sciences, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Center for Innovation in Immunoregulative Technology and Therapeutics, Graduate School of Medicine, Kyoto University, Yoshidakonoe-Cho, Sakyo-Ku, Kyoto 606-8501, Japan; Hyogo College of Medicine, 1-1 Mukogawa, Nishinomiya, Hyogo 663-8501, Japan. Electronic address:

The adoptive transfer of immune effector cells, such as CD8 killer αβ T cells, γδ T cells, NK (natural killer) cells, and genetically-modified T cells, has been receiving increasing attention. It is essential to determine cellular cytotoxicity so as to monitor the function and quality of ex vivo-expanded immune effector cells before infusion. The most common method is the [Cr]-sodium chromate release assay. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183039
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http://dx.doi.org/10.1016/j.jim.2019.01.003DOI Listing
March 2019
2 Reads

Development of a modified yeast display system for screening antigen-specific variable lymphocyte receptor B in hagfish (Eptatretus burgeri).

J Immunol Methods 2019 Mar 3;466:24-31. Epub 2019 Jan 3.

Laboratory of Aquatic Animal Diseases, Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju, Gyeongnam 660-701, South Korea. Electronic address:

The variable lymphocyte receptor B (VLRB) of jawless vertebrates has a similar function to the antibodies produced by jawed vertebrates, and has been considered as an alternative source to mammalian antibodies for use in biological research. We developed a modified yeast display vector system (pYD8) to display recombinant hagfish VLRB proteins on the extracellular surface of yeast for the isolation of antigen-specific VLRBs. After observing an up-regulation in the VLRB response in hagfish immunized with hemagglutinin 1 of avian influenza virus H9N2 subtype (H9N2-HA1), the antigen-specific VLRBs decorated on the yeast's surface were selected by quantitative library screening through magnetic-activated cell sorting (MACS) and fluorescent-activated cell sorting (FACS). Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183039
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http://dx.doi.org/10.1016/j.jim.2019.01.001DOI Listing
March 2019
11 Reads

Measurement of IL-21 in human serum and plasma using ultrasensitive MSD S-PLEX® and Quanterix SiMoA methodologies.

J Immunol Methods 2019 Mar 24;466:9-16. Epub 2018 Dec 24.

Immunology Discovery, Eli Lilly and Company, Indianapolis, IN 46285, USA.

IL-21 is a pleiotropic cytokine that plays a key role in modulating inflammatory responses, including the promotion of autoimmune diseases. Several groups have quantitated circulating levels of IL-21 in plasma and serum samples using various commercial ELISAs. We determined, however, that the most commonly used commercial assays in published literature were not specific or sensitive enough to detect levels of IL-21 in heparin plasma or serum from healthy human individuals. Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.005DOI Listing
March 2019
4 Reads

Optimization of single-cell plate sorting for high throughput sequencing applications.

J Immunol Methods 2019 Mar 24;466:17-23. Epub 2018 Dec 24.

Stanford University, Department of Medicine/Nephrology, Palo Alto, CA 94304, United States; VA Palo Alto Health Care System, Palo Alto, CA 94304, United States. Electronic address:

Single cell sequencing has recently been applied to many immunological studies. Flow cytometric index sorting isolates cells for single cell sequencing with protein level data linked to sequences. However, successful sequencing of index sorted samples requires careful optimization of several sort parameters, including nozzle size, flow rate, threshold rate, and yield calculations. Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6363834PMC
March 2019
2 Reads

Phage-based assay for rapid detection of bacterial pathogens in blood by Raman spectroscopy.

J Immunol Methods 2019 Feb 12;465:45-52. Epub 2018 Dec 12.

Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale F. Stagno d'Alcontres 31, 98166 Messina, Italy. Electronic address:

Sepsis is a systemic inflammatory response ensuing from presence and persistence of microorganisms in the bloodstream. The possibility to identify them at low concentrations may improve the problem of human health and therapeutic outcomes. So, sensitive and rapid diagnostic systems are essential to evaluate bacterial infections during the time, also reducing the cost. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183038
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http://dx.doi.org/10.1016/j.jim.2018.12.004DOI Listing
February 2019
11 Reads

An optimized protocol for the analysis of house dust mite (Dermatophagoides farinae)-induced neutrophil-dominant airway inflammation.

J Immunol Methods 2019 Feb 12;465:53-60. Epub 2018 Dec 12.

Department of Immunology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan; AMED-CREST, AMED, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. Electronic address:

House dust mites (HDMs), Dermatophagoides sp., are one of the most widespread aeroallergens worldwide and cause various allergic diseases, including asthma. The pathophysiology of asthma has been intensively investigated using murine models of allergic airway inflammation induced by exposure to D. Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.003DOI Listing
February 2019
1 Read

The relevance of buffer system ionic strength in immunoassay development.

Authors:
Axel Petzold

J Immunol Methods 2019 Feb 11;465:27-30. Epub 2018 Dec 11.

Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London WC1N 3BG, United Kingdom. Electronic address:

The best validated immunoassays for neurodegeneration have been developed for class III and IV intermediate filaments. There are a number of unique biochemical features of the intrinsically unstructured polyampholytic tail regions of these proteins which affect domain structure and thereby affinity and epitope recognition of antibodies used in immunoassays. Here one of these intermediate filaments, the neurofilament heavy chain, is chosen to demonstrate the effect of the ionic strength of a buffer system on the analytical signal to noise ratio. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183035
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http://dx.doi.org/10.1016/j.jim.2018.11.013DOI Listing
February 2019
12 Reads

Simple, rapid and inexpensive typing of common HLA class I alleles for immunological studies.

J Immunol Methods 2019 Feb 8;465:72-76. Epub 2018 Dec 8.

University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, Woolloongabba, Queensland, Australia. Electronic address:

Current HLA-typing methods are typically designed to provide exquisitely-detailed identification of multiple HLA-alleles to satisfy the requirements for organ and bone marrow transplantation or genetic studies. Many human immunological studies, on the other hand, focus around only a small number of HLA alleles that are abundant or of relevance to specific diseases. Consequently, for such studies, many HLA typing approaches are not cost-effective and are potentially complicated, slow and not easily performed in-house. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183020
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http://dx.doi.org/10.1016/j.jim.2018.12.002DOI Listing
February 2019
11 Reads

High-sensitivity lateral flow immunoassay with a fluorescent lanthanide nanoparticle label.

J Immunol Methods 2019 Feb 4;465:39-44. Epub 2018 Dec 4.

Department of Biotechnology, University of Turku, Turku, Finland.

Lateral flow (LF) immunoassays are commonly used for point-of-care testing and typically incorporate visually read reporters, such as gold particles. To improve sensitivity and develop quantitative LF immunoassays, visual reporters can be replaced by fluorescent reporters detected by an instrument. In this study, we used fluorescent europium(III) chelate doped nanoparticle (Eu-np) reporters to develop a quantitative high-sensitivity LF immunoassay for free prostate specific antigen (fPSA). Read More

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http://dx.doi.org/10.1016/j.jim.2018.12.001DOI Listing
February 2019
1 Read

Production of a recombinant monoclonal antibody to Herpes Simplex Virus glycoprotein D for immunoaffinity purification of tagged proteins.

J Immunol Methods 2019 Feb 28;465:31-38. Epub 2018 Nov 28.

Department of Biomolecular Engineering, The University of California at Santa Cruz, Santa Cruz, CA, USA. Electronic address:

We have developed a stable Chinese Hamster Ovary (CHO) cell line for the production of a recombinant monoclonal antibody (mAb) to a short protein sequence derived from the N-terminus of human herpes simplex virus type 1 glycoprotein D (HSV-1 gD). The antibody (designated r34.1) provides a useful tool for the immunoaffinity purification of HSV-1 gD tagged proteins, and provides a generic purification system by which various proteins and peptides can be purified. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.015DOI Listing
February 2019
11 Reads

Cross-species reactive monoclonal antibodies against the extracellular domains of the insulin receptor and IGF1 receptor.

J Immunol Methods 2019 Feb 27;465:20-26. Epub 2018 Nov 27.

Global Drug Discovery, Novo Nordisk, Måløv, Denmark. Electronic address:

Translation across species of immunoassay results is often challenging due to the lack of cross-species reactivity of antibodies. In order to investigate the biology of insulin and IGF1 receptors, we generated new versatile monoclonal assay antibodies using the extracellular domain of the insulin/IGF1 hybrid receptor as the bait protein in the Adimab yeast antibody discovery platform and as the antigen in a rabbit monoclonal antibody platform. The resulting antibody clones were screened for receptor specificity as well as cross-species reactivity to both tissue and cell line derived samples. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183028
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http://dx.doi.org/10.1016/j.jim.2018.11.014DOI Listing
February 2019
5 Reads

IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality.

J Immunol Methods 2019 Feb 26;465:13-19. Epub 2018 Nov 26.

Integrated Biobank of Luxembourg, 1 rue Louis Rech, 3555 Dudelange, Luxembourg; International Society for Biological and Environmental Repositories (ISBER), Biospecimen Science Working Group, Canada. Electronic address:

Background: Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs).

Methods: PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.012DOI Listing
February 2019
1 Read

A primer set for comprehensive amplification of V-genes from rhesus macaque origin based on repertoire sequencing.

J Immunol Methods 2019 Feb 22;465:67-71. Epub 2018 Nov 22.

Bioengineering Faculty of Engineering, Bar-Ilan University, Ramat Gan, Israel.

Recombinant antibodies serve as therapeutic molecules for a broad range of applications. High affinity antibodies are typically isolated following an active and effective immunization. Human-like antibodies may be obtained from immunized nonhuman primates (NHP), such as rhesus macaque, when immunized human origin is not available. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.011DOI Listing
February 2019
1 Read

Improving TCR affinity on 293T cells.

J Immunol Methods 2019 Mar 22;466:1-8. Epub 2018 Nov 22.

Division of Immune Response, Aichi, Cancer Center Research Institute, Nagoya 464-8681, Japan; Division of Cellular Oncology, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan. Electronic address:

This study presents an efficient method to improve TCR affinity, comprising 1) CDR-directed saturation mutation of TCR cDNA, 2) transient TCR display on CD3-expressing HEK293T (CD3-293T) cells by simple plasmid transfection, 3) staining with HLA-tetramers, and 4) multi-round sorting of cells with CD8-independent tetramer binding on a flow cytometer. Using these procedures, we successfully identified mutant TCRs with enhanced binding from an HLA-A*24:02-restricted, human telomerase reverse transcriptase (hTERT)-specific TCR. Two such clones, 2A7A and 2D162, harboring mutations in CDR1 and CDR2 of TCRβ, respectively, were isolated with both showing sequential four amino acid substitutions. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.010DOI Listing
March 2019
11 Reads

Unravelling enhancement of antibody fragment stability - Role of format structure and cysteine modification.

J Immunol Methods 2019 Jan 22;464:57-63. Epub 2018 Nov 22.

School of Biotechnology, Dublin City University, Dublin 9 D09 Y5N0, Ireland,; Qatar Foundation, Hamad Bin Khalifa University, Education City, Doha, Qatar.

Antibody-based diagnostics and therapeutics have huge commercial value. However, applications of antibodies are often limited by instability, particularly for recombinant antibody formats. This paper describes the conversion of a single-chain variable fragment (scFv) antibody to a single-chain antibody fragment (scAb) with notably improved stability characteristics. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.012DOI Listing
January 2019
2 Reads

Non-invasive dynamic monitoring initiation and growth of pancreatic tumor in the LSL-Kras;LSL-Trp53;Pdx-1-Cre (KPC) transgenic mouse model.

J Immunol Methods 2019 Feb 20;465:1-6. Epub 2018 Nov 20.

Department of Radiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA; Robert H. Lurie Comprehensive Cancer Center, Chicago, IL 60611, USA. Electronic address:

The LSL-Kras;LSL-Trp53;Pdx-1-Cre (KPC) mouse is one of the most widely used transgenic models to evaluate tumor characteristics and to develop novel therapies for pancreatic ductal adenocarcinoma (PDAC). There is no report of the effective systemic evaluation of longitudinal KPC tumor imitation and growth. Therefore, we aimed to characterize the initiation and progression of pancreatic cancer in KPC mice using longitudinal multiparametric magnetic resonance imaging (MRI) approaches and overall survival. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183038
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http://dx.doi.org/10.1016/j.jim.2018.11.009DOI Listing
February 2019
10 Reads

Comparison of bead array and glass nanoreactor multi-analyte platforms for the evaluation of CNS and peripheral inflammatory markers during HIV infection.

J Immunol Methods 2019 Feb 20;465:7-12. Epub 2018 Nov 20.

Department of Medicine, University of California at San Diego School of Medicine, La Jolla, CA, United States; Department of Psychiatry, University of California at San Diego School of Medicine, La Jolla, CA, United States.

While human immunodeficiency virus (HIV) infection has become a treatable disease with the development of combination antiretroviral therapy (cART), chronic inflammation that affects the central nervous system and other organs is still common. Reliable methods are needed to study HIV-associated inflammatory biomarkers. In this study involving both plasma and cerebrospinal fluid (CSF), we compared multiplex bead array (MBA) to a relatively new technology based on microfluidics and glass nanoreactor (GNR) technology for the measurement of three commonly studied markers from HIV-infected individuals. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.008DOI Listing
February 2019
8 Reads

Updates in diagnostic and clinical laboratory immunology from the 30th annual meeting of the Association of Medical Laboratory Immunologists (AMLI).

J Immunol Methods 2019 Jan 16;464:138-140. Epub 2018 Nov 16.

Pediatrics and Pathology, Keck School of Medicine, University of Southern California, Laboratory Medicine, Children's Hospital of Los Angeles, CA, USA.

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http://dx.doi.org/10.1016/j.jim.2018.11.007DOI Listing
January 2019
2 Reads

Effect of peripheral blood mononuclear cell cryopreservation on innate and adaptive immune responses.

J Immunol Methods 2019 Feb 14;465:61-66. Epub 2018 Nov 14.

Pneumococcal Research, Murdoch Children's Research Institute, Melbourne, Melbourne, VIC 3052, Australia; Department of Paediatrics, University of Melbourne, Australia. Electronic address:

Cryopreservation of blood-derived immune cells is commonly used in clinical trials to examine immunological responses. However, studies elucidating the effects of cryopreservation on peripheral blood mononuclear cell (PBMC) responses have shown inconsistent results making it difficult to draw meaningful conclusions. Therefore we sought to address this issue by comparing key innate and adaptive immune parameters between freshly-isolated and cryopreserved PBMCs from healthy adults. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183030
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http://dx.doi.org/10.1016/j.jim.2018.11.006DOI Listing
February 2019
10 Reads

Isolation of tumor endothelial cells from murine cancer.

J Immunol Methods 2019 Jan 3;464:105-113. Epub 2018 Nov 3.

Department of Gastroenterological and Transplant Surgery, Institute of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima 734-8551, Japan. Electronic address:

Tumor endothelial cells (TECs), which constitute the lining of the tumor blood vessels, have various characteristics as tumor constituent cells. In this study, we describe a novel method for the isolation of highly pure, fresh TECs, which form a small population within the tumor. Tumors were first dissected from tumor-bearing mice and digested to a single cell suspension with Collagenase Type II; the single cells were then separated by density gradient centrifugation. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.005DOI Listing
January 2019
1 Read

Secreted and intracellular cytokines are complementary measures for human monocytes treated with Toll-like receptor agonists.

J Immunol Methods 2019 Jan 3;464:131-137. Epub 2018 Nov 3.

Department of Medicine, University of Melbourne, Melbourne, Australia; Immunology Research Centre, St. Vincent's Hospital, Melbourne, VIC, Australia. Electronic address:

Cytokine production by human peripheral blood mononuclear cells including monocytes, is frequently assessed by measuring secreted cytokines using enzyme linked immunosorbent assay (ELISA), whereby the total concentration of one cytokine of interest is obtained without information regarding the cell type responsible for making the cytokine. Cytokines can be retained inside the cell using protein transport inhibitors. Subsequent analysis by flow cytometry not only identifies the cell type producing the cytokine but can semi-quantitate the amount of cytokine produced by measuring the geometric mean fluorescence intensity (gMFI) and is amenable to analyzing more than one protein associated with the same cell (multiplexing). Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183022
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http://dx.doi.org/10.1016/j.jim.2018.11.003DOI Listing
January 2019
6 Reads
2.005 Impact Factor

CLEC4C gene expression can be used to quantify circulating plasmacytoid dendritic cells.

J Immunol Methods 2019 Jan 3;464:126-130. Epub 2018 Nov 3.

The University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, Brisbane, QLD, Australia; Department of Respiratory Medicine, Princess Alexandra Hospital, Woolloongabba, QLD, Australia. Electronic address:

Plasmacytoid dendritic cells (pDC) are an important type I interferon producer that play an important role in the first line of host defence during viral infection. Abnormalities in pDC numbers and function have been associated with several health conditions. Quantifying pDC is important for understanding pDC related immune responses in viral infections and other diseases, however the current methods for quantifying pDC using flow cytometry have limited utility in large cohort studies involving multiple centres with limited access to flow cytometry. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183033
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http://dx.doi.org/10.1016/j.jim.2018.11.001DOI Listing
January 2019
10 Reads

Validation of an ADCC assay using human primary natural killer cells to evaluate biotherapeutic products bearing an Fc region.

J Immunol Methods 2019 Jan 2;464:87-94. Epub 2018 Nov 2.

Unidad de Desarrollo e Investigación en Bioprocesos, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de México, Mexico. Electronic address:

The development of biotherapeutics requires continuous improvement in analytical methodologies for the assessment of their quality attributes. A subset of biotherapeutics is designed to interact with specific antigens that are exposed on the membranes of target cells or circulating in a soluble form, and effector functions are achieved via recognition of their Fc region by effector cells that induce mechanisms such as antibody-dependent cell-mediated cytotoxicity (ADCC). Thus, ADCC induction is a critical quality attribute (CQA) that must be evaluated to ensure biotherapeutic efficacy. Read More

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http://dx.doi.org/10.1016/j.jim.2018.11.002DOI Listing
January 2019
2 Reads

Development and validation of a mouse-based primary screening method for testing relative allergenicity of proteins from different wheat genotypes.

J Immunol Methods 2019 Jan 3;464:95-104. Epub 2018 Nov 3.

Food Allergy & Immunology Laboratory, Michigan State University, East Lansing, MI 48824, United States. Electronic address:

Background: Wheat allergy is a major food allergy that has reached significant levels of global public health concern. Potential variation in allergenicity among different wheat genotypes is not well studied at present largely due to the unavailability of validated methods. Here, we developed and validated a novel mouse-based primary screening method for this purpose. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183032
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http://dx.doi.org/10.1016/j.jim.2018.11.004DOI Listing
January 2019
10 Reads
2.005 Impact Factor

Development of a simple new flow cytometric antibody-dependent cellular cytotoxicity (ADCC) assay with excellent sensitivity.

J Immunol Methods 2019 Jan 31;464:74-86. Epub 2018 Oct 31.

Drug Discovery Antibody Platform Unit, RIKEN Center for Integrative Medical Science (IMS), 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan. Electronic address:

Antibody-based therapeutic strategies have become recognized as useful clinical options in several types of cancer, often with the expectation that such therapies will trigger target cell elimination via antibody-dependent cellar cytotoxicity (ADCC) by natural killer cells. The successful development of therapeutic monoclonal antibodies (mAbs) requires an assay system that permits a critical evaluation of their physicochemical and biological characteristics. At present a number of ADCC assay systems have been reported, however, there is still room for improvement in terms of usability, operability and sensitivity. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.014DOI Listing
January 2019
1 Read

A high throughput lentivirus sieving assay identifies neutralization resistant Envelope sequences and predicts in vivo sieving.

J Immunol Methods 2019 Jan 31;464:64-73. Epub 2018 Oct 31.

ImmunoTechnology Section, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892, United States. Electronic address:

An effective prophylactic vaccine against human immunodeficiency virus (HIV) will likely require a potent antibody response that can neutralize the virus at the mucosal portal of entry. The elicitation of potent broadly-neutralizing anti-sera will be an iterative process, optimizing candidates that only block a fraction of potential viral strains. This effect, termed "sieving", is evidence of a partially efficacious vaccine. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6322957PMC
January 2019
1 Read

Application of the immunoregulatory receptor LILRB1 as a crystallisation chaperone for human class I MHC complexes.

J Immunol Methods 2019 Jan 24;464:47-56. Epub 2018 Oct 24.

Cancer Immunology and Immunotherapy Centre, Institute of Immunology and Immunotherapy, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK. Electronic address:

X-ray crystallographic studies of class I peptide-MHC molecules (pMHC) continue to provide important insights into immune recognition, however their success depends on generation of diffraction-quality crystals, which remains a significant challenge. While protein engineering techniques such as surface-entropy reduction and lysine methylation have proven utility in facilitating and/or improving protein crystallisation, they risk affecting the conformation and biochemistry of the class I MHC antigen binding groove. An attractive alternative is the use of noncovalent crystallisation chaperones, however these have not been developed for pMHC. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183025
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http://dx.doi.org/10.1016/j.jim.2018.10.011DOI Listing
January 2019
12 Reads

Expression and characterization of soluble epitope-defined major histocompatibility complex (MHC) from stable eukaryotic cell lines.

J Immunol Methods 2019 Jan 19;464:22-30. Epub 2018 Oct 19.

Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Sciences Center, Abilene, TX 79601, United States. Electronic address:

MHC class I-specific reagents such as fluorescently-labeled multimers (e.g., tetramers) have greatly advanced the understanding of CD8+ T cells under normal and diseased states. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183034
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http://dx.doi.org/10.1016/j.jim.2018.10.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6322931PMC
January 2019
9 Reads

Immunofluorescence staining of live lymph node tissue slices.

J Immunol Methods 2019 Jan 19;464:119-125. Epub 2018 Oct 19.

Department of Chemistry, University of Virginia, Charlottesville, USA; Department of Biomedical Engineering, University of Virginia, Charlottesville, USA; Carter Immunology Center, University of Virginia, Charlottesville, USA. Electronic address:

Explants of lymphoid tissue provide a rare opportunity to assess the organization of the immune system in a living, dynamic environment. Traditionally, ex vivo immunostaining is conducted in fixed tissue sections, while live tissues are analyzed using genetically engineered fluorescent reporters or adoptively transferred, pre-labelled cell populations. Here, we validated a protocol for immunostaining and imaging in live, thick slices of lymph node tissue, thus providing a spatial "map" of the lymph node while maintaining the viability and functionality of the slices. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183028
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http://dx.doi.org/10.1016/j.jim.2018.10.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6322934PMC
January 2019
9 Reads

The influence of EDTA Vacutainer blood collection tube on the level of blood interleukin-1 receptor antagonist.

J Immunol Methods 2019 Jan 19;464:114-118. Epub 2018 Oct 19.

Department of Epidemiology, UCLA, Fielding School of Public Health, Los Angeles, CA 90095, USA; Department of Medicine, UCLA, David Geffen School of Medicine, Los Angeles, CA 90095, USA.

Background: The use of anticoagulants may influence the composition of blood cells and interfere with plasma levels of IL-1ra when unprocessed EDTA blood samples are stored for long periods of time.

Methods: Blood was drawn into EDTA and heparinized blood collection tubes from 11 HIV-1 negative men participating in the Multicenter AIDS Cohort Study (MACS) and 4 healthy volunteers. The blood was processed according to the experiments detailed in the method and after incubation; supernatants were collected and stored at -70 °C until batch testing using IL-1ra ELISA. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.009DOI Listing
January 2019
1 Read

Mitochondrial membrane potential identifies cells with high recombinant protein productivity.

J Immunol Methods 2019 Jan 17;464:31-39. Epub 2018 Oct 17.

Cell Culture and Fermentation Sciences, MedImmune, Gaithersburg, MD, United States.

Development of cell lines for biotherapeutic protein production requires screening large numbers of clones to identify and isolate high producing ones. As such, stable cell line generation is a time- and resource-intensive process. There is an increasing need to enhance the selection efficiency of high-yielding clonal cell lines for cell line development projects by using high throughput screening of live cells for markers predictive of productivity. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.007DOI Listing
January 2019
1 Read

Analytical validation of an alternative method to quantify specific antibodies in 3 applications.

J Immunol Methods 2019 Jan 18;464:40-46. Epub 2018 Oct 18.

Paris Descartes University, Paris, France; Hôpital Européen Georges Pompidou, APHP, Service d'Immunologie Biologique, Paris, France; Centre de Recherche des Cordeliers, Université Pierre et Marie Curie, INSERM UMRS1138, Paris, France. Electronic address:

The detection and the quantification of specific antibodies represent essential tools for the diagnosis and for the biological monitoring of immune humoral response in many clinical situations in particular in autoimmune diseases or in the context of immunotherapy using monoclonal antibodies. This article focuses on the development of a specific antibody measuring method (Patent n°PCT/IB2014/064437). The principle of this method is based on the combined use of a monoclonal antibody as standard and the protein G as immunoglobulins detecting agent. Read More

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http://dx.doi.org/10.1016/j.jim.2018.10.008DOI Listing
January 2019
2 Reads

Detection of activated neutrophils by reactive oxygen species production using a hematology analyzer.

J Immunol Methods 2018 Dec 16;463:122-126. Epub 2018 Oct 16.

Kobe University Hospital, Kobe, Japan.

Neutrophils are recruited to infection sites and kill bacteria by phagocytosis and reactive oxygen species (ROS) production. It has been reported that vacuoles are present in neutrophils that produce ROS and are present in large numbers in blood smears of patients with bacterial infections. The leukocyte differentiation function on the Sysmex automated hematology analyzer classifies leukocytes by flow cytometry. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183006
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http://dx.doi.org/10.1016/j.jim.2018.10.004DOI Listing
December 2018
12 Reads

Enhancing recombinant antibody performance by optimally engineering its format.

J Immunol Methods 2018 Dec 12;463:127-133. Epub 2018 Oct 12.

School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland; Hamad Bin Khalifa University and Research, Development and Innovation, Qatar Foundation, Doha, Qatar.

Antibody-based sensors are now widely used in therapeutics, diagnostics, and in environmental monitoring. Recombinant antibodies are becoming integral parts of such devices due to their reported high affinities, their capacity for engineering to achieve highly defined performance characteristics and the fact that their production can be optimized to a significant degree. To aid as a model for the identification of important analyte binding residues within the antibody sub-structure and elucidate the docking characteristics of small molecules such as metabolites, illicit drugs, biotherapeutics (proteins, peptides and nucleic acids) or toxins towards the antibody, herein we report the binding of the harmful cyanobacterial-toxin, microcystin-leucine-arginine (MC-LR) to a single chain fragment variable (scFv) antibody fragment. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183033
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http://dx.doi.org/10.1016/j.jim.2018.10.005DOI Listing
December 2018
10 Reads

An optimized protocol to quantify signaling in human transitional B cells by phospho flow cytometry.

J Immunol Methods 2018 Dec 13;463:112-121. Epub 2018 Oct 13.

Division of Nephrology, Department of Medicine, The University of Illinois at Chicago, 820 South Wood Street (MC 793), Chicago, IL 60612, USA.

Background And Purpose: Phospho flow cytometry is a powerful technique to analyze signaling in rare cell populations. This technique, however, requires harsh conditions for cell fixation and permeabilization, which can denature surface antigens or antibody-conjugated fluorochromes. These are among several technical limitations which have been a barrier to quantify signaling in unique B cell subsets. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183017
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http://dx.doi.org/10.1016/j.jim.2018.10.002DOI Listing
December 2018
6 Reads

Sequencing the peripheral blood B and T cell repertoire - Quantifying robustness and limitations.

J Immunol Methods 2018 Dec 10;463:137-147. Epub 2018 Oct 10.

Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, United States. Electronic address:

The adaptive immune response generates a large repertoire of T cells with T-cell receptors (TCR and TCR) and B cells with immunoglobulins (Ig). The repertoire changes in response to antigen stimulation both through amplification of specific cells (clonal expansion) as well as somatic hypermutation of immunoglobulins. Alterations of the immune repertoire have been observed in response to acute disease, such as external pathogens, or chronic diseases, such as autoimmunity and cancer. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183030
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http://dx.doi.org/10.1016/j.jim.2018.10.003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6355145PMC
December 2018
11 Reads

Excessive outlier removal may result in cut points that are not suitable for immunogenicity assessments.

J Immunol Methods 2018 Dec 9;463:105-111. Epub 2018 Oct 9.

MedImmune LLC, One MedImmune Way, Gaithersburg, MD 20878, United States.

Cut point determination is an important aspect of immunogenicity assay development. The cut point can be influenced by a myriad of factors. Key among those is the analytical variability of the assay itself and biological variation due to test samples. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759183027
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http://dx.doi.org/10.1016/j.jim.2018.10.001DOI Listing
December 2018
5 Reads

The role of the laboratory in the expanding field of neuroimmunology: Autoantibodies to neural targets.

Authors:
Stanley J Naides

J Immunol Methods 2018 Dec 6;463:1-20. Epub 2018 Oct 6.

Immunology R&D, Quest Diagnostics Nichols Institute, 33608 Ortega Highway, San Juan Capistrano, CA 92675, USA. Electronic address:

Accelerated identification of autoantibodies associated with previously idiopathic neurological disease has provided insights into disease mechanisms, enhanced understanding of neurological function, and opportunities for improved therapeutic interventions. The role of the laboratory in the expanding field of neuroimmunology is critical as specific autoantibody identification provides guidance to clinicians in diagnosis, prognosis, tumor search strategies, and therapeutic interventions. The number of specific autoantibodies identified continues to increase and newer testing strategies increase efficiencies in the laboratory and availability to clinicians. Read More

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https://linkinghub.elsevier.com/retrieve/pii/S00221759173048
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http://dx.doi.org/10.1016/j.jim.2018.04.003DOI Listing
December 2018
2 Reads

Comparison of gene expression and flow cytometry for immune profiling in chronic lymphocytic leukaemia.

J Immunol Methods 2018 Dec 26;463:97-104. Epub 2018 Sep 26.

ACRF Translational Research Laboratory, Royal Melbourne Hospital, Melbourne, Victoria, Australia; Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Melbourne, Victoria, Australia.

Understanding how cancer and cancer therapies affect the immune system is integral to the rational application of immunotherapies. Flow cytometry is the gold standard method of peripheral blood immune cell profiling. However, the requirement for viable cells can limit its applicability, especially in studies of retrospective clinical cohorts. Read More

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http://dx.doi.org/10.1016/j.jim.2018.09.013DOI Listing
December 2018
2 Reads